Applications for pulmonary biospecimens include, but are not limited to analyses of immune cell trafficking and signal transduction as well as inflammatory mediator expression.

Collection, storage, and assessment:

Bronchoalveolar lavage (BAL) samples. Collected BAL samples are centrifuged to pellet the cellular fraction. Cell are counted and differentiated on cytospin preps using Wight-Geimsa stain. Upon request, cell differentials are then determined from at least 500 leukocytes using standard hematological criteria. Remaining cells are stored as cell pellets at -80oC for RNA, DNA, or protein analyses. Supernatants are stored at -80oC until distribution.

Plasma samples. Collected blood samples are ficolled and cells are harvested from buffy coat layers. Cells are then counted and differentiated on cytospin preps using Wright-Geimsa stain. Upon request, cell differentials are then determined from at least 500 leukocytes using standard hematological criteria. Remaining cells are stored as cell pellets at -80C for RNA, DNA, or protein analyses. Plasma fluid samples are stored at -80oC until distribution.

Serum samples. Collected blood samples are centrifuged and the serum is aliquoted and stored at -80oC until distribution.

Sample organization and quality control. All samples are logged into the PBR using FreezerWorks, a state-of-the-art cataloging program designed for the organization of stored clinical samples. Cell and fluid samples are assessed routinely for viability. Specifically, cells are lysed and assayed for ƒÒ-actin mRNA content via RT-PCR; fluids are examined for TNFƒÑ and/or TGFƒÒ protein levels via ELISA.

Patient clinical history. Each sample is linked directly with the respective patient clinical history via P-TREC, a HIPPA compliant comprehensive clinical database developed by the Division of Pulmonary, Allergy and Critical Care Medicine and hosted by the Division of Preventive Medicine.